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Old 05-20-2011, 05:00 AM   #1
vepached64
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Location: State college

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Default Mixed sample and starting amount questions

Hello all,
I know RNA seq using SOLiD can be done from RNA isolated from a single cell which is in nanogram quanitity.

Now, we have a sample of DNA (circular plus linear) whose size ranges from 10 kb to 6 MB, isolated from some where. I would like to sequence it completely, like genomic DNA sequencing.

Question 1: How much DNA is needed to do it by either 454 sequencing or SOLiD sequencing?
Do we really need microgram or milligram quantities of DNA? If so, what is the use of advanced techs, for processing an archaeological sample?

Question 2: Can we do Exome sequencing of an unknown sample?
Any help is greatly appreciated.

Question 3: Any suggestions as best approach for complete sequencing or find out ORFs in the sample/

Venkata

Last edited by vepached64; 05-20-2011 at 05:02 AM. Reason: One more question
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Old 05-20-2011, 10:04 AM   #2
ECO
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You need to do some searching and reading on your own. Moving this to general as it's not related per se to SOLiD
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Old 05-30-2011, 06:46 AM   #3
zhengz
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Hi,

To answer a little part of your Q1, one nanogram after fragmentation was enough for 10 454 Titanium runs.

To just test what was in a mixed sample, sub-picogram DNA was used for a mini-454 sequencing (Sanger sequencing using 454 library on a 96-well plate):

http://dx.plos.org/10.1371/journal.pone.0019723.
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